pRS314-hUD
(Plasmid #11005)

• Depositing Lab: Francisco Malagon
• Publication: Unpublished, Paco (unpublished)

Backbone

• Vector backbone: pRS314
• Backbone size w/o insert (bp): 4783
• Vector type: Bacterial Expression, Yeast Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: hUD recombination system
• Alt name: his3.3'D::URA3::his3.5'D
• Species: S. cerevisiae (budding yeast)
• Insert Size (bp): 3047
• Mutation: Direct repeat recombination system. Recombination reconstitutes a functional HIS3 gene

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: SacI (not destroyed)
• 3′ cloning site: XhoI (not destroyed)
• 5′ sequencing primer: T7
• 3′ sequencing primer: T3

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

The repeats of HIS3 in this plasmid contain truncations of the HIS3 gene. The first repeat is a 3' end truncation that contains the promoter and the ORF from the ATG to the internal KpnI site (used for subcloning). The second repeat is a 5' truncation that contains from the internal NdeI site (used for subcloning) to the end of the gene. Recombination of the common DNA sequence between these repeats (approx. 430 nt internal to the HIS3 ORF) reconstitutes a functional gene. Both plasmids, containing the repeats in direct orientation (hUD) or inverted orientation (hUI), have been tested for recombination with positive result.